Hydrogel pullulan nanoparticles encapsulating pBUDLacZ plasmid as an efficient gene delivery carrier

Area of Science:

• Biotechnology
• Nanomedicine
• Cell Biology

Background:

• Effective nucleic acid delivery into cells is crucial for gene therapy and research.
• Developing safe and efficient delivery vehicles remains a significant challenge in biotechnology.
• Hydrogel nanoparticles offer potential as novel drug and gene delivery systems.

Purpose of the Study:

• To develop and characterize pullulan nanoparticles for enhanced nucleic acid delivery.
• To evaluate the safety and efficacy of pullulan nanoparticles in vitro.
• To compare the transfection efficiency of pullulan nanoparticles with a commercial delivery agent.

Main Methods:

• Pullulan nanoparticles encapsulating pBUDLacZ plasmid were prepared using w/o microemulsions.
• Particle size and morphology were analyzed using Transmission Electron Microscopy (TEM).
• Cytotoxicity was assessed via MTT assay, and cellular uptake was visualized using Scanning Electron Microscopy (SEM).
• Transfection efficiency was evaluated in HEK293 and COS-7 cells.

Main Results:

• Spherical pullulan nanoparticles with a diameter of 45 nm were successfully prepared.
• Nanoparticles exhibited excellent biocompatibility, with over 100% cell viability at concentrations up to 1000 microg/ml.
• SEM confirmed nanoparticle internalization, leading to vacuole formation and cytoskeleton disruption (F-actin, beta-tubulin).
• Transfection efficiency was cell-type dependent, with COS-7 cells showing higher beta-galactosidase expression compared to HEK293 cells.
• Beta-gal expression in COS-7 cells was comparable to Lipofectamine 2000.

Conclusions:

• Pullulan nanoparticles are a safe and effective system for intracellular nucleic acid delivery.
• The observed transfection efficiency in COS-7 cells suggests potential for gene therapy applications.
• Further development of pullulan nanoparticles could lead to advanced drug and gene delivery platforms.